Results:Membranefractionsfromthree-dayand200mMsalt-treatedArabidopsissuspensionplantswereisolated,followedbyproteaseshavingandenrichmentusingZirconiumion-chargedmagneticb... Results: Membrane fractions from three-day and 200 mM salt-treated Arabidopsis suspension
plants were isolated, followed by protease shaving and enrichment using Zirconium ion-charged
magnetic beads, and tandem mass spectrometry analyses. From this isolation, 18 phosphorylation
sites from 15 Arabidopsis proteins were identified. A unique phosphorylation site in 14-3-3-
interacting protein AHA1 was predominately identified in 200 mM salt-treated plants. We also
identified some phosphorylation sites in aquaporins. A doubly phosphorylated peptide of PIP2;1 as
well as a phosphopeptide containing a single phosphorylation site (Ser-283) and a phosphopeptide
containing another site (Ser-286) of aquaporin PIP2;4 were identified respectively. These two sites
appeared to be novel of which were not reported before. In addition, quantitative analyses of
protein phosphorylation with either label-free or stable-isotope labeling were also employed in this
study. The results indicated that level of phosphopeptides on five membrane proteins such as
AHA1, STP1, Patellin-2, probable inactive receptor kinase (At3g02880), and probable purine
permease 18 showed at least two-fold increase in comparison to control in response to 200 mM
salt-stress. 展开
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